title: Comparison of fluorescent labels for oligosaccharides and introduction of a new postlabeling purification method.
abstract: Labeling of oligosaccharides with fluorescent dyes is the prerequisite for their sensitive analysis by high-performance liquid chromatography (HPLC). In this work, we present a fast new postlabeling cleanup procedure that requires no device other than the reaction vial itself. The procedure can be applied to essentially all labeling reagents. We also compare the performance of 15 different labels for N-glycan analysis in various analytical procedures. We took special care to prevent obscuring influences from incomplete derivatization and signal quenching by impurities. Procainamide emerged as more sensitive than anthranilic acid for normal-phase HPLC, but its chromatographic performance was not convincing. 2-aminopyridine was the label with the lowest retention on reversed-phase and graphitic carbon columns and, thus, appears to be most suitable for glycan fractionation by multidimensional HPLC. Most glycan derivatives performed better than native sugars in matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and electrospray ionization-MS (ESI-MS), but the gain was small and hardly sufficient to compensate for sample loss during preparation.
authors.affiliation: Department of Chemistry, University of Natural Resources and Applied Life Sciences (BOKU), 1190 Vienna, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Pabst
authors.forename: Daniel
authors.initials: D
authors.lastname: Kolarich
authors.forename: Gerald
authors.initials: G
authors.lastname: Pöltl
authors.forename: Thomas
authors.initials: T
authors.lastname: Dalik
authors.forename: Gert
authors.initials: G
authors.lastname: Lubec
authors.forename: Andreas
authors.initials: A
authors.lastname: Hofinger
authors.forename: Friedrich
authors.initials: F
authors.lastname: Altmann
mesh_terms: D000096:Acetone / Q000737:chemistry; D000586:Amination; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000737:chemistry*; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D011134:Polysaccharides / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D005456:Fluorescent Dyes; D009844:Oligosaccharides; D011134:Polysaccharides; D000096:Acetone